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Visualizing the enteric nervous system using genetically engineered double reporter mice: Comparison with

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  • 1Division of Gastroenterology, Department of Medicine, School of Medicine, University of California, San Diego, California.

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Summary

We developed a novel genetically engineered mouse to visualize enteric nervous system (ENS) neurons expressing neuronal nitric oxide synthase (nNOS) and choline acetyltransferase (ChAT). This reporter mouse allows for efficient study of ENS in gastrointestinal diseases.

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Area of Science:

  • Neuroscience
  • Gastroenterology

Background:

  • The enteric nervous system (ENS) regulates gastrointestinal functions.
  • Traditional immunohistochemistry is used to visualize ENS neurons.
  • Genetically engineered mice offer an alternative for neuron visualization.

Purpose of the Study:

  • To develop a mouse model for visualizing enteric nervous system (ENS) neurons.
  • To express fluorescent neuronal nitric oxide synthase (nNOS) and choline acetyltransferase (ChAT) in ENS neurons.
  • To compare visualization in reporter mice versus traditional immunostaining.

Main Methods:

  • Generated a reporter mouse line using Cre/LoxP technique for nNOS expression.
  • Crossbred mice hemizygous for ChAT-ChR2-YFP BAC transgene with nNOS reporter mice.
  • Examined tissues using fluorescent microscopy and compared with wild-type immunostaining.

Main Results:

  • Over 95% of ENS neurons expressed either nNOS or ChAT or both.
  • nNOS and ChAT neurons were well visualized throughout the GI tract in reporter mice.
  • Fluorescence localization differed, with nNOS seen in cytoplasm and nucleus in reporter mice versus cytoplasm only in immunostained samples.

Conclusions:

  • The developed double reporter mouse is a powerful tool for studying the ENS.
  • This model facilitates research on the effects of diseases on the ENS.
  • It offers an alternative to traditional immunostaining methods for ENS research.